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WO2008065540A3 - Method for the detection of post-translational modifications - Google Patents

Method for the detection of post-translational modifications Download PDF

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Publication number
WO2008065540A3
WO2008065540A3 PCT/IB2007/004341 IB2007004341W WO2008065540A3 WO 2008065540 A3 WO2008065540 A3 WO 2008065540A3 IB 2007004341 W IB2007004341 W IB 2007004341W WO 2008065540 A3 WO2008065540 A3 WO 2008065540A3
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WO
WIPO (PCT)
Prior art keywords
post
protein substrate
translational modification
cells
binding
Prior art date
Application number
PCT/IB2007/004341
Other languages
French (fr)
Other versions
WO2008065540A2 (en
Inventor
Eric Trinquet
Achim Brinker
Emmanuel Claret
Gerard Mathis
Original Assignee
Cis Bio Int
Eric Trinquet
Achim Brinker
Emmanuel Claret
Gerard Mathis
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cis Bio Int, Eric Trinquet, Achim Brinker, Emmanuel Claret, Gerard Mathis filed Critical Cis Bio Int
Priority to EP07859356.3A priority Critical patent/EP2087355B1/en
Priority to JP2009537717A priority patent/JP2010510771A/en
Priority to US12/516,333 priority patent/US8663928B2/en
Publication of WO2008065540A2 publication Critical patent/WO2008065540A2/en
Publication of WO2008065540A3 publication Critical patent/WO2008065540A3/en

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
    • G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
    • C12Q1/485—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase involving kinase

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Method for the detection in homogeneous medium of a post-translational modification of a protein substrate catalyzed by a cell enzyme, characterized in that the post-translational modification reaction takes place in intact living cells, in that these cells comprise a heterologous expression vector coding for a fusion protein comprising the protein substrate and a first coupling domain and in that it comprises the following stages: (v) Incubation of the cells in the presence or in the absence of a compound to be tested capable of modulating the activity of said enzyme, (vi) Addition to the reaction medium of a first fluorescent compound member of a first pair of FRET partners covalently bonded to a coupling agent capable of binding specifically to the first coupling domain present on the protein substrate, (vii) Addition to the reaction medium of a second fluorescent compound member of this first pair of FRET partners, and covalently bonded to a binding domain specific to the site of the protein substrate having undergone the post-translational modification and not binding to the non-modified protein substrate, (i) Measurement of the FRET signal emitted by the sample, this signal being representative of the quantity of protein substrate having undergone said post-translational modification; and cells for the implementation of said method.
PCT/IB2007/004341 2006-11-27 2007-11-27 Method for the detection of post-translational modifications WO2008065540A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP07859356.3A EP2087355B1 (en) 2006-11-27 2007-11-27 Method for the detection of post-translational modifications
JP2009537717A JP2010510771A (en) 2006-11-27 2007-11-27 Methods for detection of post-translational modifications
US12/516,333 US8663928B2 (en) 2006-11-27 2007-11-27 Method for the detection of post-translational modifications

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US86108506P 2006-11-27 2006-11-27
US60/861,085 2006-11-27

Publications (2)

Publication Number Publication Date
WO2008065540A2 WO2008065540A2 (en) 2008-06-05
WO2008065540A3 true WO2008065540A3 (en) 2008-10-23

Family

ID=39468330

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2007/004341 WO2008065540A2 (en) 2006-11-27 2007-11-27 Method for the detection of post-translational modifications

Country Status (4)

Country Link
US (1) US8663928B2 (en)
EP (1) EP2087355B1 (en)
JP (1) JP2010510771A (en)
WO (1) WO2008065540A2 (en)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5954728B2 (en) * 2010-01-22 2016-07-20 国立研究開発法人科学技術振興機構 Probe reagent for measuring protein degradation activity
CN103630691B (en) * 2012-08-24 2015-07-08 中国科学院深圳先进技术研究院 Bioassays for small ubiquitin-like modifications
CA2897340C (en) 2013-01-09 2022-09-06 Friedrich-Alexander-Universitat Erlangen-Nuernberg Method for in vitro detection and monitoring of a disease by measuring disease-associated protease activity in extracellular vesicles
CN104749143A (en) * 2013-12-31 2015-07-01 深圳先进技术研究院 Detection method for sumoylated modification of proteins and application thereof
KR101750407B1 (en) 2014-11-10 2017-06-27 한양대학교 산학협력단 Phosphorylated Peptide-Specific RNA Aptamer
CN107463795A (en) * 2017-08-02 2017-12-12 南昌大学 A kind of prediction algorithm for identifying tyrosine posttranslational modification site
US11604186B2 (en) * 2018-10-17 2023-03-14 Molecular Devices (Austria) GmbH Real time western blot assays utilizing fluorescence resonance energy transfer (FRET)
EP4433824A1 (en) * 2021-11-16 2024-09-25 Biogen MA Inc. Method for detecting a lipidated protein via fluorescence resonance energy transfer (fret)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001036617A2 (en) * 1999-11-18 2001-05-25 Cyclacel Limited Assay for measuring enzyme activity in vivo

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6900304B2 (en) 1996-01-31 2005-05-31 The Regents Of The University Of California Emission ratiometric indicators of phosphorylation
GB9901072D0 (en) * 1999-01-19 1999-03-10 Imp Cancer Res Tech Methods for detecting changes to a macromolecular component of a cell
JP2004537047A (en) * 2001-06-14 2004-12-09 アナディーズ　ファーマスーティカルズ，アイエヌシー． Methods for screening ligands for target molecules
CA2445420A1 (en) * 2003-07-29 2005-01-29 Invitrogen Corporation Kinase and phosphatase assays
US20060194262A1 (en) * 2004-10-15 2006-08-31 Shuichan Xu P27 ubiquitination assay and methods of use

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001036617A2 (en) * 1999-11-18 2001-05-25 Cyclacel Limited Assay for measuring enzyme activity in vivo

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
GAUTIER I ET AL: "Homo-FRET microscopy in living cells to measure monomer-dimer transition of GFP-tagged proteins", BIOPHYSICAL JOURNAL, NEW YORK, US, US, vol. 80, no. 6, 1 June 2001 (2001-06-01), pages 3000 - 3008, XP002977865, ISSN: 0006-3495 *
NI ET AL: "Analyzing protein kinase dynamics in living cells with FRET reporters", METHODS : A COMPANION TO METHODS IN ENZYMOLOGY, ACADEMIC PRESS INC., NEW YORK, NY, US, vol. 40, no. 3, 1 November 2006 (2006-11-01), pages 279 - 286, XP005716995, ISSN: 1046-2023 *

Also Published As

Publication number Publication date
WO2008065540A2 (en) 2008-06-05
US20100086943A1 (en) 2010-04-08
EP2087355A2 (en) 2009-08-12
JP2010510771A (en) 2010-04-08
US8663928B2 (en) 2014-03-04
EP2087355B1 (en) 2014-09-24

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